Histomorphological Parameters Research Articles

Comparison of Del Nido and histidine-tryptophan-ketoglutarate cardioplegia solutions: an animal study with prolonged ischaemia

ObjectiveMyocardial protection is important for a successful procedure cardiac surgery, and the key element of myocardial protection is cardioplegia. We compared Del Nido cardioplegia (DN) and Bretschneider histidine-tryptophan-ketoglutarate cardioplegia (HTK) regarding cardioprotective effects in a porcine model of prolonged ischaemia.MethodsLandrace pigs weighing 50–60 kg were randomized to receive either DN (n = 9) or HTK (n = 9). All pigs underwent cardiac arrest for 90 min followed by 120 min of reperfusion/convalescence. A detailed set of laboratory, histological and functional parameters was acquired at baseline, during cardiac arrest and following reperfusion/convalescence.ResultsPressure-volume measurements revealed better systolic and diastolic left ventricular performance in DN as compared to HTK (both p < 0.05). Haemoglobin decreased after application of the cardioplegic solution. The decrease was more pronounced in the HTK group than in the DN group (p < 0.01). In contrast to DN, sodium (p < 0.01) and chloride levels (p < 0.05) were significantly decreased in the HTK group after initiation of CPB and remained decreased after reperfusion. The number of animals requiring defibrillations to restore sinus rhythm significantly differed between the groups [HTK: 100% (n = 9/9) vs. DN: 44.4% (n = 4/9), p = 0.03]. Expression of ICAM-1 as a marker of endothelial dysfunction was lower in the DN group compared to the HTK group (p = 0.02). Histological evaluation, oxidative and nitrosative stress, mitochondrial membrane integrity and apoptosis markers were comparable between DN and HTK groups (all p > 0.05).ConclusionsIn this porcine model with prolonged ischaemia, DN was superior to HTK in terms of haemoglobin levels, blood electrolytes, spontaneous return of sinus rhythm, left ventricular function, and endothelial injury. Histomorphological parameters indicative of ischaemia/reperfusion injury, oxidative stress and mitochondrial function as well as apoptosis-inducing factors did not differ.

Study of histomorphological changes in the subarachnoid hemorrhage at different time intervals between injury infliction and death.

Even though biochemical and molecular methods are under research for wound dating purpose, histology of the injury is still the mainstay of wound dating technique. Significance of knowing how old the wound is can have major implication in the delivery of justice. Similarly, subarachnoid hemorrhage may need to be dated in cases where other data for wound dating are scarce. This study is designed to study the histomorphological changes in the subarachnoid hemorrhages of different time interval since injury and aim to use this information for dating of subarachnoid hemorrhage for medicolegal purpose. The histology samples of subarachnoid hemorrhage of the deceased with head injuries with known post injury interval were taken during the autopsies and divided into different groups. The gross color changes and histomorphological parameters were evaluated in each group and statistical inference was made. The result showed gross color, RBC lysis, macrophage infiltration, hemosiderin laden macrophage, fibrin deposition, collagen deposition and meningeal reactive changes significantly correlated with post injury interval. Whereas, Neutrophils, lymphocytes and phagocytosis did not correlate. RBC lysis also correlated with gross color change as well. In conclusion, histology of subarachnoid hemorrhage can be a useful tool in dating subarachnoid hemorrhage in cases where it is required.

Impact of Chronic Noise Stress on Thyroid Health: A Comparative Histomorphological and Endocrine Study in Adult Male and Female Rats.

To compare the effect of varying durations of noise exposure on the histomorphological and endocrine profile of the thyroid gland in male and female adult rats. An experimental study. Place and Duration of the Study: Department of Anatomy, Army Medical College, Rawalpindi / NUMS, from January to December 2020. Thirty Sprague-Dawley rats (15 males and 15 females) were divided into three groups each having 10 rats with equal male- to-female ratio. Group A was the control group. Group B and Group C were exposed to 100 dB noise for 4 hours daily and 100 dB noise for 6 hours daily for 4 months, respectively. Thyroid histomorphological parameters such as follicular epithelial cells' height, follicular diameter, and thyroid stimulating hormone (TSH) levels were evaluated at the end of the study. Both male and female rats exposed to noise stress exhibited reduced follicular diameter (p <0.001) and increased epithelial cells' height (p <0.001) as compared to the control group. The experimental groups showed significantly higher TSH levels compared to the control group (p <0.001) among female rats, while no significant difference was observed among male rats (p 0.47). No significant differences in histological and serum parameters were noted between the experimental groups (Group B and C). Chronic noise exposure induces hyperactivity of thyroid follicles in both male and female adult rats as evidenced by the thyroid histology, whereas only female rats showed raised TSH levels, suggesting a potential disruption in the hypothalamic-pituitary-thyroid axis. Thyroid gland, Chronic noise stress, Follicular epithelial cells, Thyroid stimulating hormone, Thyroid follicles.

Impact of high-fructose diet and metformin on histomorphological and molecular parameters of reproductive organs and vaginal microbiota of female rat

There are limited data on the effects of a high-fructose diet on the female reproductive system. Although metformin has some functional effects on female fertility, its reproductive outcome on high fructose diet-induced metabolic syndrome is unclear. The aim of the present study is to evaluate the impact of a high fructose diet on histomorphological and molecular parameters of the reproductive organs and vaginal microbiota as well as the treatment potential of metformin. Wistar albino rats were used in the study. The metabolic syndrome model was induced by a high-fructose diet in rats for 15 weeks. Metformin was orally administered once a day for the last 6 weeks. The high-fructose diet increased blood glucose, triglycerides, insulin, and ovarian testosterone levels; however, it reduced ovarian aromatase levels and follicle numbers and caused uterine inflammation. The high-fructose diet-induced molecular abnormalities on ovarian tissue were demonstrated by the downregulation of ovarian insulin signaling pathway proteins and dysregulation of ovarian mitogenic and apoptotic pathway proteins. A high-fructose diet caused vaginal dysbiosis, metformin increased probiotic bacteria in the vaginal microbiota. Our results revealed that metformin improves ovarian impairments by modulating hormonal balance, insulin level, mapk, and apoptotic signaling molecules, as well as regulating the vaginal microbiota.

Relevance of the Immunohistochemical Expression of p53 and E-cadherin in the Grading of Urothelial Carcinoma: A Single-Center Cross-Sectional Observational Study.

Background Urothelial carcinoma (UC) is a prevalent cancer worldwide, primarily affecting the urinary bladder. It is more common in men than women and is often linked to factors like tobacco smoking, occupational exposure, and chronic infections. UC can be classified into different subtypes based on its growth pattern (papillary or non-papillary) and the extent of invasion into the bladder wall. The management of UC depends on its grade and stage, with treatment options ranging from transurethral resection of bladder tumor (TURBT) for non-invasive tumors to cystectomy for muscle-invasive disease. This study aimed to investigate the expression of p53 and E-cadherin in low-grade and high-grade UC and their correlation with histomorphological parameters. Materials and methods A cross-sectional observational study was conducted on 50 histopathologically confirmed UC cases. Immunohistochemical (IHC) staining for p53 and E-cadherin was performed, and the results were correlated with clinicopathological features. Statistical analysis was performed to find the correlation between p53 and E-cadherin immunoexpression and the grade of UC. Results The cohort's mean age was 58.18±12.99 years, with a male predominance (76%). Most cases (52%) were from rural areas and unskilled workers (52%). Hematuria was present in all cases (100%), while urgency and frequency were reported in 32% each. High-grade UC was more common (76%), with 48% being invasive. Muscle invasion was absent in 56%. p53 overexpression was seen in all cases, with 38% moderate and 36% strong staining. Twenty-two out of 24 cases (91.7%) of invasive UC showed low E-cadherin expression, while all 26 cases (100%) of low-grade papillary UC and high-grade papillary UC displayed high E-cadherin expression. This suggests that reduced E-cadherin expression is strongly associated with invasive UC, potentially as a tumor aggressiveness and progression marker. Conclusion Considering that elevated p53 protein expression is linked to aggressive tumor behavior, a more intensive treatment strategy is recommended for patients with non-muscle-invasive bladder cancer (NMIBC) who show high p53 protein scores. On the other hand, the downregulation or absence of E-cadherin expression has been recognized as a strong indicator of advanced grade and higher clinical stages.

The prognostic significance of growth pattern, tumor budding, poorly differentiated clusters, desmoplastic reaction pattern and tumor-stroma ratio in colorectal cancer and an evaluation of their relationship with KRAS, NRAS, BRAF mutations

Growth pattern (GP), tumor budding (TB), poorly differentiated clusters (PDC), desmoplastic reaction pattern (DRP) and tumor-stroma ratio (TSR) are prognostic histomorphological parameters in colorectal cancer (CRC). Correlations between these parameters, their individual prognostic values, and their relationship with KRAS/NRAS/BRAF mutations have not been comprehensively examined. We aimed to investigate these associations, which have not been previously explored in this combination. 126 CRC cases were included. GP, TB, PDC, DRP and TSR were evaluated by two experienced pathologists. KRAS/NRAS/BRAF mutation profile were determined using qPCR. Demographic, clinicopathological and survival data were recorded. Interrelations were investigated by statistical analysis. Infiltrative GP was more frequent in high-score TB, PDC-G3, and stroma-high tumors (p < 0.05). High-score TB was more common in PDC-G3 and stroma-high tumors (p < 0.05). Immature DRP was more frequent in stroma-high tumors (p = 0.014). Among histomorphological parameters, a significant relationship was found only between infiltrative GP and the presence of KRAS mutation (p = 0.023). Moreover, GP was significantly associated with pT, lymphatic invasion, perineural invasion (p < 0.05). Effects on survival were assessed using Kaplan-Meier method and Cox proportional hazards model. TB and PDC were identified as independent predictors of overall survival. Higher TB score (p = 0.008) and higher PDC grade (p = 0.013) lead to worse survival. Interestingly, GP, DRP, TSR or KRAS/NRAS/BRAF mutations were not associated with overall survival. Our results highlight the prognostic significance of TB and PDC. We suggest incorporating TB and PDC into routine CRC reports. The association of KRAS mutation with infiltrative GP supports its role in the acquisition of invasive behavior.

Histological and Biochemical Alterations Underlying Statin-Associated Pancreatic Toxicity in a Rat Model

ObjectiveTo investigate the histological and biochemical alterations in the pancreas associated with statin-induced toxicity using a rat model. MethodologyA one-year laboratory-based experimental control trial conducted at department of anatomy of a public sector medical college in Rawalpindi, in collaboration with the National Institute of Health (NIH) Islamabad and Armed Forces Institute of Pathology (AFIP) Rawalpindi. Twenty male Sprague-Dawley rats were assigned to the control group and another twenty to the experimental group. For a duration of 12 weeks, the experimental group was given 60 mg/kg/day of simvastatin orally, whereas the control group was given water and a conventional diet. The following characteristics were measured: body weight, pancreatic weight, biochemical parameters (cytokines, enzymes), and histological alterations (congestion, necrosis, fibrosis). Version 23 of the Statistical Package for the Social Sciences (SPSS) program was used to analyze the data. Qualitative variables were expressed as percentages and frequencies. The statistical significance was defined as P &lt; 0.05. Chi-square analyses contrasted the qualitative factors among the groups. ResultsCompared to controls, the simvastatin group showed significantly increased mean pancreatic weight (p=0.043), congestion (55% vs 0%, p&lt;0.001), higher levels of CRP (p&lt;0.030), IL-6 (p&lt;0.040), and TNF-α (p&lt;0.040). Trends of mild necrosis (25% vs 5%, p=0.182) and interstitial fibrosis (15% vs 0%, p=0.231) were observed but not statistically significant. DiscussionThis work shows that simvastatin significantly affects the pancreatic histomorphology and biochemical parameters in rats. It causes a noticeable amount of necrosis, interstitial fibrosis, and changes the pancreatic biochemical profile. These changes raise the possibility of long-term statin-associated pancreatitis and endocrine/exocrine dysfunction. Using a stable animal model, these new findings offer distinctive insights into the impact of simvastatin on pancreatic morphology. Conclusion Simvastatin profoundly impacts pancreatic histomorphology, warranting further investigation into mechanisms of toxicity and close monitoring of patients for pancreatic adverse effects during statin therapy.

Developmental variations of the reproductive organs of ganders from different goose breeds and the underlying mechanisms

A deep understanding of the dynamics and mechanisms of male reproductive tract development is necessary for adoption of either genetic techniques or environmental management practices for improving fertility and hatchability in poultry. However, compared with other poultry such as chickens and ducks, less is known about the age- and breed-related changes in the reproductive tract development of domestic goose ganders exhibiting relatively poor reproductive performance as well as the regulatory mechanisms. In the present study, by taking 2 Chinese domestic goose breeds (Sichuan White goose, SW and Gang goose, GE; Anser cygnoides) and one European goose breed (Landes goose, LD; Anser anser) as the experimental objects, we comprehensive analyzed the morphological, histological, and genome-wide transcriptomic variations in their testicular and external genital development during the period from hatching to sexual maturity. Results from histomorphological analysis demonstrated that the reproductive tract of all goose breeds developed in both age- and breed-dependent manners, and the left and right testis developed asymmetrically throughout posthatch development. The tenth week posthatch was a critical developmental stage for all goose ganders, because both the testicular and external genital histomorphological parameters significantly changed before and after this period. During the first 10 wk posthatch, the weight, organ index, or size of male reproductive organs developed more rapidly in SW than in LD, and so were the testicular parenchymal-to-interstitial ratio and the external genital lymphatic lumen diameter. However, the testicular seminiferous epithelium thickness, seminiferous tubule diameter, and Leydig cell number, as well as the external genital keratinized epithelium thickness were significantly higher in LD than in SW at 10 wk of age. Through comparative transcriptomics analysis and RT-qPCR validation, several pathways related to germ and somatic cell function, organ remodeling, and energy metabolism were thought to be responsible for the developmental variations in the early testicular development between Chinese and European domestic ganders, where 10 hub genes involved in the cell cycle, RNA polymerase II-dependent transcription, and mitotic cell division pathways might play essential roles. These data shed new light on the interbreed differences in the male goose reproductive tract development and the molecular mechanisms regulating male goose testicular functions and fertility.

Perinatal Exposure to Glyphosate or a Commercial Formulation Alters Uterine Mechanistic Pathways Associated with Implantation Failure in Rats.

Perinatal exposure to a glyphosate-based herbicide (GBH) or its active ingredient, glyphosate (Gly), has been demonstrated to increase implantation failure in rats. This study investigates potential mechanisms of action, analyzing uterine preparation towards the receptive state. Pregnant Wistar rats (F0) were treated orally with GBH or Gly (3.8 and 3.9 mg Gly/kg/day, respectively) from gestational day (GD) 9 until weaning. Adult F1 females became pregnant and uterine samples were collected on GD5 (preimplantation period). Histomorphological uterine parameters were assessed. Immunohistochemistry was applied to evaluate cell proliferation and protein expression of estrogen receptors (ERα and ERβ), cell cycle regulators (PTEN, cyclin G1, p27, and IGF1R-α), and the Wnt5a/β-catenin/FOXA2/Lif pathway. Both GBH and Gly females showed increased stromal proliferation, associated with a high expression of ERs. Dysregulation of PTEN and cyclin G1 was also observed in the Gly group. Reduced gland number was observed in both groups, along with decreased expression of Wnt5a/β-catenin/FOXA2/Lif pathway in the glandular epithelium. Overall, GBH and Gly perinatal exposure disrupted intrinsic uterine pathways involved in endometrial proliferation and glandular function, providing a plausible mechanism for glyphosate-induced implantation failure by compromising uterine receptivity. Similar effects between GBH and Gly suggest the active principle mainly drives the adverse outcomes.

Renal Protective Effect of Boeravinone B against Diabetic Nephropathy Rats via Inhibition of The Inflammatory and JAK2/STAT3 Signalling Pathway.

Chronic inflammation is a common feature in diabetes, especially when blood sugar levels are poorly controlled. This chronic low-grade inflammation can affect various organs, including the kidneys. Podocyte damage play a key role in the development of diabetic nephropathy (DN). The aim of the study was to evaluate the nephroprotective effect of Boeravinone B (BB) against streptozotocin (STZ) induced DN in rats and explore the underlying mechanism. In this experimental study, the rats received intraperitoneal injections of STZ (60 mg/kg) to induce DN. Various doses of BB (2.5, 5, and 7.5 mg/kg) were administered orally. Glucose levels, body weights, and organ weights (hepatic and renal) were assessed. Renal, histomorphological, antioxidant, hepatic, and cytokine levels were determined, as were the mRNA expression levels of JAK2 and STAT3. At end of the experimental study, the rats were sacrificed and their renal tissues were removed for histopathological assessment. BB treatment decreased glucose levels and increased body weights. This treatment suppressed hepatic weights, increased renal tissue weights, and also decreased renal parameters like uric acid, urea, bilirubin, creatinine (Cr) and, albumin. There was a decrease (P<0.001) in histomorphological parameters such as kidney hypertrophy index (KHI), mean glomerular volume (MGV), foot process fusion ratio (FPFR), and glomerular basement membrane thickness (GBMT) after treatment with BB. In addition, this treatment improved the levels of renal podocin, renal CD2- associated protein (CD2AP) and suppressed hepatic parameter levels. BB treatment (P<0.001) altered antioxidant parameters and cytokine levels, and suppressed mRNA expressions of JAK2, STAT3, RAGE, KIM-1, NAGL, and S100A8. Administration of BB showed renal protective effects against STZ-induced DN in rats via the reduction of oxidative stress and inflammatory reactions.

Multicenter Study on Tumor Budding in Lung Squamous Cell Carcinoma: Comparison Between Biopsy and Resection With Interobserver Variability Assessment

Grading lung squamous cell carcinoma (LUSC) is controversial and not universally accepted. The histomorphologic feature of tumor budding (TB) is an established independent prognostic factor in colorectal cancer, and its importance is growing in other solid cancers, making it a candidate for inclusion in tumor grading schemes. We aimed to compare TB between preoperative biopsies and resection specimens in pulmonary squamous cell carcinoma and assess interobserver variability. A retrospective cohort of 249 consecutive patients primarily resected with LUSC in Bern (2000-2013, n = 136) and Lausanne (2005-2020, n = 113) with available preoperative biopsies was analyzed for TB and additional histomorphologic parameters, such as spread through airspaces and desmoplasia, by 2 expert pathologists (M.M., C.N.). Results were correlated with clinicopathologic parameters and survival. In resection specimens, peritumoral budding (PTB) score was low (0-4 buds/0.785 mm2) in 47.6%, intermediate (5-9 buds/0.785 mm2) in 27.4%, and high (≥10 buds/0.785 mm2) in 25% of cases (median bud count, 5; IQR, 0-26). Both the absolute number of buds and TB score were similar when comparing tumor edge and intratumoral zone (P = .192) but significantly different from the score obtained in the biopsy (P < .001). Interobserver variability was moderate, regardless of score location (Cohen kappa, 0.59). The discrepant cases were reassessed, and consensus was reached in all cases with identification of causes of discordance. TB score was significantly associated with stage (P = .002), presence of lymph node (P = .033), and distant metastases (P = .020), without significant correlation with overall survival, tumor size, or pleural invasion. Desmoplasia was significantly associated with higher PTB (P < .001). Spread through airspaces was present in 34% and associated with lower PTB (P < .001). To conclude, despite confirming TB as a reproducible factor in LUSC, we disclose areas of scoring ambiguity. Preoperative biopsy evaluation was insufficient in establishing the final TB score of the resected tumor.

Effects of a supraphysiological dose of testosterone cypionate on salivary gland function in adult male Wistar rats

The abusive use of anabolic androgenic steroids has become a serious health problem worldwide, but its effects on oral health are still poorly understood. Therefore, the objective of this study was to evaluate the effects of a supraphysiological dose of testosterone cypionate (TC) on salivary biochemical, histomorphology, immunohistochemistry, and redox state parameters of parotid and submandibular glands. Twenty male Wistar rats, 12 weeks old, were divided into two groups (n=10/group): a control group and TC group, which received a dose of 20 mg/kg, once a week, for 6 weeks. Post treatment, the saliva and glands were collected. A supraphysiological dose of TC increased plasma and salivary testosterone concentrations. Although TC did not alter salivary flow, pH, and buffering capacity, the treatment increased the salivary secretion of total protein and reduced amylase, calcium, phosphate, and potassium. TC reduced the connective tissue area in the parotid gland and acinar area of the submandibular gland, while increasing the granular convoluted tubule area in the submandibular gland. Proliferating cell nuclear antigen was higher in the acinar cells of the submandibular glands from the TC group. Moreover, TC increased concentrations of total oxidant capacity and damaged lipids in both salivary glands, while total antioxidant activity and uric acid were lower in the submandibular gland, and reduced glutathione was higher in both glands. Superoxide dismutase, catalase, and glutathione peroxidase activities were higher in the parotid gland, while only glutathione peroxidase activity was lower in the submandibular gland of the TC group. In conclusion, TC abuse may be a potential factor for dysfunction of the parotid and submandibular glands, becoming a risk factor for the oral and systemic health of users.

Morphometric features of the duodenal wall in piglets during different periods of postnatal and neonatal ontogenesis under the influence of the ‘Globigen Jump Start’ feed additive

The morphological parameters of the small intestinal mucosa, such as villi size and crypts, play a key role in the formation of the intestinal tube’s absorption surface. The number of goblet and enterochromaffin cells indicates the epithelium’s functional condition in terms of mucus secretion and production of catecholamines and hormones (serotonin, dopamine). That is why it is important to study and determine certain morphometric parameters of the duodenal wall in piglets during different periods of neonatal and postnatal development, especially during stress situations, namely weaning and transition to protein feeding. The article presents the resulting effect studies of ‘Globigen Jump Start’ feed additive on the histo-morphological parameters of the piglets’ duodenal mucosa on day 7, 14 and 28 of life. A positive effect on mitigating weaning stress was manifested by a decrease in the quantitative and linear indicators of histoarchitectonics of intestinal wall’s individual morphological components. A significant increase in goblet cells, especially in the experimental group, had a positive impact on the mucopolysaccharide synthesis. The piglets’ gastrointestinal tract physiology involves a complex interaction between the central nervous system, metasympathetic nervous system, APUD system, and endocrine system. Due to these systems, the information is transmitted according to the direct and reverse communication mechanisms in the regulation of the gastrointestinal tract function. An increase in the number of enterochromaffin cells and their nuclei diameter in the experimental group of piglets indicated an increased synthesis of catecholamines and hormones. It has an extremely positive effect on the immune and physiological status of piglets, thus emphasizing the crucial role of serotonin in neuronal metabolism and the formation of stress resistance. The increase in the thickness of the duodenal wall muscle layer, in our opinion, occured due to the active peristalsis, which was enhanced by the action of some catecholamines, which were synthesized in a slightly larger amount under the influence of nutrients making up the ‘Globigen Jump Start’ feed additive.

Evaluation of the Expression of Programmed Death-Ligand 1 and Its Role in Differentiating Low-Grade and High-Grade Urothelial Carcinoma.

Urothelial carcinoma (UC) is a common malignancy, predominantly affecting males. Many tumor cells use the interaction between programmed death-ligand 1 (PD-L1) and programmed death receptor (PD-1) to inactivate T-cells in the microenvironment and evade host immune response. Our study aims to evaluate the expression of PD-L1 in UC and correlate its expression with histomorphological parameters. After obtaining approval from the Institute Ethics Committee, we conducted a prospective observational study on transurethral resection of urinary bladder tumor (TURBT) and cystectomy specimens histopathologically diagnosed as UC between 2022 and 2023, comprising 50 cases. All standard protocol was followed and immunohistochemistry (IHC) was done using PD-L1 with rabbit anti-human PD-L1 monoclonal antibody (Clone: IHC411; Biogenics Inc., San Francisco, CA, USA). Results: Among the 50 cases of UC, the majority were papillary type (35 cases), high grade (28 cases), and non-muscle invasive (30 cases). Among the cases studied, 15 of them showed PD-L1 positivity; 55% of the cases of muscle-invasive bladder cancer were found to be positive for PD-L1 out of which the results were statistically significant. PD-L1 expression by IHC staining can differentiate between muscle-invasive and non-muscle-invasive UC cases. This observation allows for further exploring the potential role of immune checkpoint inhibitors in adjuvant and neoadjuvant therapy, especially in muscle-invasive cases of UC.

Pathologic assessment of hepatocellular carcinoma in the era of immunotherapy: a narrative review.

Immune checkpoint inhibitor (ICI)-based therapy has achieved impressive success in various cancer types. Several ICIs have been unprecedentedly approved as the treatment regimens for advanced hepatocellular carcinoma (HCC) in recent decade. Meanwhile, numerous clinical trials are being performed to exploit more ICIs into initially unresectable HCC and postoperative HCC to expectantly induce adequate tumor downstaging for further resection or implement adjuvant treatment for relapse-free survival, respectively. In this review, we aim to summarize some pragmatic histomorphologic, immunohistochemical, and molecular pathologic parameters which promisingly indicate the response of neoadjuvant/conversion ICI-related therapy and predict the efficacy of adjuvant/therapeutic ICI-related therapy for HCC. We searched PubMed using the terms hepatocellular carcinoma, immunotherapy, immune checkpoint inhibitor, immune checkpoint blockade, conversion therapy, neoadjuvant therapy, adjuvant therapy, biomarker, pathologic evaluation, pathologic assessment till February 2023. Although there is no consensus regarding the pathologic evaluation of relevant HCC specimens, it is encouraging that a few of studies have concentrated on this field, and moreover, the methods and parameters noted on other cancer types are also worthy of reference. For the pathologic assessment of HCC specimens underwent immunotherapy, a suitable sampling scheme, identifying immunotherapy-related pathologic response, and quantification of pathologic response rate should be emphasized. For the patients of HCC who are scheduled to receive immunotherapy, tumor-infiltrating lymphocyte, intratumoral tertiary lymphoid structure, programmed cell death ligand 1, Wnt/β-catenin, microsatellite instability and mismatch repair, tumor mutational burden and tumor neoantigen, as well as some other signaling pathways are the potential predictive biomarkers of treatment response of ICI. The management of HCC in the era of immunotherapy arises a brand-new pathological challenge that is to provide an immunotherapy-related diagnostic report. Albeit many related researches are preclinical or insufficient, they may tremendously alter the immunotherapy strategy of HCC in future.

Effects of Commercial Probiotics on the Growth Performance, Intestinal Microbiota and Intestinal Histomorphology of Nile Tilapia (Oreochromis niloticus) Reared in Biofloc Technology (BFT).

Though different types of commercial probiotics are supplemented in biofloc technology (BFT), very little information is available on their effects on the farmed fish. Therefore, this study focused on evaluating the effects of three most commonly used commercial probiotics on the growth performance, intestinal histomorphology, and intestinal microbiota of Nile tilapia (Oreochromis niloticus) reared in BFT. Tilapia fry, with an average weight of 3.02 ± 0.50 g, were stocked at a density of 60 fry/0.2 m3, and cultured for 90 days. Three commercial probiotics were administered, with three replications for each: a single-genus multi-species probiotic (Bacillus spp.) (T1), a multi-genus multi-species probiotic (Bacillus sp., Lactobacillus sp., Nitrosomonas sp., Nitrobacter sp.) (T2), and a multi-species probiotic (Bacillus spp.) combined with enzymes including amylase, protease, cellulase, and xylanase (T3). The results showed significant variations in growth and feed utilization, with T3 outperforming other treatments in terms of weight gain, liver weight, and intestine weight. Adding Bacillus spp. with enzymes (T3) to water significantly increased the histomorphological parameters (villi length, villi depth, crypt depth, muscle thickness, intestinal thickness) as well as microbes (total viable count and total lactic acid bacteria) of intestine of fish compared to T1 and T2, leading to improved digestion and absorption responses. It is concluded that the supplementation of commercial probiotics has potential benefits on farmed fish species in BFT.

Evaluation of Ovarian Histomorphology and Function Following Clomiphene Citrate and Human Chorionic Gonadotropin Administration in Wistar Rats

Polycystic ovarian syndrome (PCOS), affecting 5-10% of women, is a leading cause of infertility affecting 10-15% of couples globally. Human chorionic gonadotropin (hCG) and clomiphene citrate (CC) are often utilized for the treatment of PCOS. Accordingly, this study explored the effects of CC and hCG on ovarian histomorphology and fertility parameters. Twenty adult female rats were divided into four groups as follows: Group A (control) received only feed and water; Group B received 0.7 mg/kg BW of CC twice daily for five days and was mated before sacrifice on day 19 (before litter); Group C received 0.7 mg/kg BW of CC twice daily for five days, followed by mating and allowed to litter before sacrifice; Group D received 0.7 mg/kg body weight (BW) of hCG on day one, followed by 0.7 mg/kg BW of CC twice daily for five days, and were mated before sacrifice on day 7 (before litter). Results showed that Group B rats had higher, more than other groups, Follicle-stimulating hormone (FSH), Estradiol, Testosterone and Prolactin levels when compared to the control. Similarly, rats in Group B had higher levels of Progesterone while Group D had higher levels of Luteinizing hormone (LH) when compared to the control group. Histological findings demonstrated diverse impacts on ovarian structures, ranging from congested blood vessels to haemorrhages and follicular cysts. Consequently, this study underscores the complexities of drug interactions in reproductive health and provides preliminary insights into the possible adverse effects of CC and hCG on the ovary and fertility parameters.

Efficacy of quercetin in ameliorating hypoxia-induced hematological and histopathological alterations in rohu Labeo rohita.

Hypoxia, a major issue in aquatic ecosystems, in special reference to climate change, and exacerbated by anthropogenic activities. It is causing slow growth, disease outbreaks, and mortality in finfish and shellfish. Therefore, adaptation to lowering oxygen levels through supplementation of herbs or their extracts in diets is imperative. In this study, hypoxia was simulated in controlled conditions with quercetin-enriched diets. Quercetin is a plant pigment (flavonol) possessing anti-oxidant property and is present in vegetables, leaves, seeds, pulses, and fruits. The experiment was conducted on rohu Labeo rohita, which is most widely cultured in India. There were four treatments including T1 (Normoxia: > 5ppm dissolved oxygen; DO2), T2 (hypoxia: 3-4ppm DO2), T3 (hypoxia + 50mg quercetin/kg diet), and T4 (hypoxia + 100mg quercetin/kg diet). The study was conducted for 30days, and water quality was measured regularly. The results revealed that the hematological parameters were negatively affected. The tissue micro-architecture illustrated the impairment through degeneration of neurons in the brain, increased pigmentation as melanosis in the kidney, increased thickness of primary lamellae in the gills, and dilatations of sinusoids in the liver in hypoxia groups, while quercetin-enriched diets improved the hematological and histomorphological parameters. The results confirm the utility of hematological and histopathological tools as biomarkers and reflect the possible threats of hypoxia on fish. In conclusion, quercetin in diets appeared to show resistance towards chronic hypoxia by restoring the structure and functions of the vital organs towards normalcy and could be recommended as a potential ameliorative agent.

Neoadjuvant Radiochemotherapy Alters the Immune and Metabolic Microenvironment in Oral Cancer-Analyses of CD68, CD163, TGF-β1, GLUT-1 and HIF-1α Expressions.

The first-line treatment of oral squamous cell carcinoma (OSCC) involves surgical tumor resection, followed by adjuvant radio(chemo)therapy (R(C)T) in advanced cases. Neoadjuvant radio- and/or chemotherapy has failed to show improved survival in OSCC. Recently, neoadjuvant immunotherapy has shown promising therapeutic efficacy in phase 2 trials. In this context, the addition of radio- and chemotherapy is being reconsidered. Therefore, a better understanding of the tumor-biologic effects of neoadjuvant RCT would be beneficial. The current study was conducted on a retrospective cohort of patients who received neoadjuvant RCT for the treatment of oral cancer. The aim of the study was to evaluate the influence of neoadjuvant RCT on the immunological tumor microenvironment (TME) and hypoxic and glucose metabolisms. A cohort of 45 OSSC tissue samples from patients were analyzed before and after RCT (total 50.4 Gy; 1.8 Gy 5× weekly; Cisplatin + 5-Fluorouracil). Immunohistochemistry for CD68, CD163, TGF-β, GLUT-1 and HIF-1α was performed using tissue microarrays and automated cell counting. Differences in expression before and after RCT and associations with histomorphological parameters (T-status, N-status) were assessed using the Mann-Whitney U test. Tumor resection specimens after neoadjuvant RCT showed a significant decrease in CD68 infiltration and a significant increase in CD163 cell density. The CD68/CD163 ratio was significantly lower after RCT, indicating a shift toward M2 polarization. The GLUT-1 and HIF-1α expressions were significantly lower after RCT. Larger tumors (T3/T4) showed a lower GLUT-1 expression. Other biomarkers were not associated with the T- and N-status. Neoadjuvant RCT with 50.4 Gy induced a shift toward the M2 polarization of macrophages in the TME. This change in immune composition is not favorable and may be prognostically negative and counteract immunotherapeutic approaches. In addition, the decreased expressions in GLUT-1 and HIF-1α indicate reductions in the glucose metabolism and hypoxic energy metabolism in response to "high dose" neoadjuvant RCT, which may be therapeutically desirable.

Chromium Picolinate Regulates Bone Metabolism and Prevents Bone Loss in Diabetic Rats.

Diabetic osteoporosis (DOP) is an abnormal metabolic disease caused by long-term hyperglycemia. In this study, a model rat of streptozotocin (STZ)-induced diabetes was established, and chromium picolinate (5 mg·kg-1) was given; the changes in blood glucose and body weight were detected before and after administration; and bone mineral density (BMD), bone morphology, bone turnover markers, inflammatory cytokines, and oxidative stress indicators were observed in each group. We found that after chromium picolinate (CP) intervention for 8 weeks, the blood glucose level was decreased; the BMD, the bone histomorphology parameters, and the pathological structure were improved; the expression of bone resorption-related proteins was downregulated; and the expression of bone formation-related proteins was upregulated. Meanwhile, serum antioxidant activity was increased, and inflammatory cytokine levels were decreased. In conclusion, CP could alleviate DOP by anti-oxidation, inhibition of bone turnover, anti-inflammation, and regulation of the OPG/RANKL/RANK signaling pathway. Therefore, CP has important application values for further development as a functional food or active medicine in DOP treatment.